Early detection of influenza A(H5) viruses with affinity for the human sialic acid receptor by MALDI-TOF mass spectrometry based mutation detection

J Virol Methods. 2011 Mar;172(1-2):72-7. doi: 10.1016/j.jviromet.2010.12.024. Epub 2010 Dec 30.

Abstract

Highly pathogenic avian influenza (HPAI) A(H5N1) strains have been causing sporadic cases of disease in South East Asia and Africa for many years. These cases are associated with a high fatality rate, and it is feared that the virus could evolve into a strain capable of causing a pandemic. It is likely that a requirement for a A(H5) pandemic to occur is a switch in the receptor affinity of the virus. Candidate mutations in the hemagglutinin glycoprotein have been identified in the literature, and their emergence in circulating viruses would be an ominous development. This study describes a method to identify the presence of these mutations, even within a quasispecies, using RT-PCR followed by in vitro translation and peptide characterization by MALDI-TOF mass spectrometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Cloning, Molecular
  • DNA, Complementary
  • Genome, Viral
  • Hemagglutinin Glycoproteins, Influenza Virus / chemistry
  • Hemagglutinin Glycoproteins, Influenza Virus / genetics
  • Hemagglutinin Glycoproteins, Influenza Virus / metabolism
  • Humans
  • Influenza A Virus, H5N1 Subtype / genetics*
  • Influenza A Virus, H5N1 Subtype / metabolism*
  • Influenza, Human / diagnosis
  • Influenza, Human / virology*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Mutation*
  • Protein Binding
  • Receptors, Cell Surface / metabolism*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization*

Substances

  • DNA, Complementary
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Receptors, Cell Surface
  • sialic acid receptor